Skip to main content
. 2018 May 10;7:e35258. doi: 10.7554/eLife.35258

Figure 1. Inducible tkv allele switch system.

(A) Schematic illustration of a FLP/FRT-mediated tkv allele switch. mCherry-tagged Tkv expression was specifically induced in the wing pouch via FLP/FRT-mediated recombination that eliminates tkv-GFP from the locus. White and grey boxes represent tkv coding and 3’UTR sequences, respectively. (B–G) tkvGFP>>mCherry heterozygous animal before and after allele switch. (B, C, E, F) GFP (green) and mCherry (red) represent endogenous Tkv expression in the developing wing disc. BMP/DPP activity was detected by anti-p-Mad staining (grey). (D, G) Both Tkv-GFP and Tkv-mCherry expressing animals developed normal adult wings. Longitudinal veins L2-L5 are indicated. (H–J) Wing pouch-specific allele switch of the constitutively active form of Tkv, TkvQ199D. TkvQ199D-mCherry (red) expression led to ectopic p-Mad activation (grey, H, I), resulting in wing malformation (J). Scale bars: 50 μm for (B), and 0.5 mm for (D). Anterior is oriented to the left in all wing disc figures and to the top in adult wing images.

Figure 1.

Figure 1—figure supplement 1. Wing patterning defects caused by tkv overexpression.

Figure 1—figure supplement 1.

(A, B) p-Mad staining of nub-Gal4/+ wing disc (A) and nub-Gal4/+ adult wing (B). (C, D) tkv overexpression in the wing pouch by nub-Gal4 (nub-Gal4/UAS-tkv) led to abnormal p-Mad expression (C), resulting in wing patterning defects (D). Scale bars: 50 μm for (A), 0.5 mm for (B). Anterior is oriented to the left in all wing disc figures and to the top in adult wing images.

Figure 1—figure supplement 2. Spatial regulation of tkv allele switch.

Figure 1—figure supplement 2.

(A) Flowchart illustrating the establishment of tkv allele switch transgenic lines. (B–E) Region-specific induction of Tkv-mCherry using the Gal4/UAS system. Control (tkvGFP>>mCherry/+, B), the posterior (tkvGFP>>mCherry/+; hh-Gal4, UAS-FLP/+, C), the dorsal (tkvGFP>>mCherry/ap-Gal4; UAS-FLP/+, D), and the wing pouch switch (tkvGFP>>mCherry/nub-Gal4; UAS-FLP/+, E). Scale bars: 100 μm. Anterior is to the left.